Characterization of Bucolome N-Glucuronide Formation: Tissue Specificity and Identification of Rat UDP-Glucuronosyltransferase Isoform (s)

DOI: 10.4236/pp.2011.23021   PDF   HTML     4,038 Downloads   7,859 Views   Citations


Bucolome N-glucuronide (BCP-NG, main metabolite of bucolome (BCP) is the first N-glucuronide of barbituric acid derivatives isolated from rat bile. The objective of this study was to identify the main tissue producing BCP-NG and the molecular species of BCP-NG-producing UGT. Four target tissues were investigated: the liver, small and large intestines, and kidney. To identify the UGT molecular species responsible for BCP-NG formation, yeast microsomes expressing each rat UGT isoform were prepared. BCP-NG formation was detected in all microsomal fractions of the 4 tissues. The liver microsomal BCP-NG-producing activity was the highest, followed by that in the small intestinal microsomes, showing about 41% of the liver microsomal activity level. BCP-NG-producing activity (min-1) was determined in yeast microsomal fractions expressing rat UGT isoforms, and the activity was detected in UGT1A1 (0.059), UGT1A2 (0.318), UGT1A3 (0.001), UGT1A7 (0.003), UGT2B1 (0.004), UGT2B3 (0.091), and UGT2B6 (0.031), showing particularly high levels for UGT1A1 and UGT1A2 among the UGT1A isoforms. It was clarified that UGT1A1, widely distributed in rat tissues, is the molecular species responsible for BCP-NG formation.

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H. Kanoh, M. Tada, S. Ikushiro and K. Mohri, "Characterization of Bucolome N-Glucuronide Formation: Tissue Specificity and Identification of Rat UDP-Glucuronosyltransferase Isoform (s)," Pharmacology & Pharmacy, Vol. 2 No. 3, 2011, pp. 151-158. doi: 10.4236/pp.2011.23021.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] S. Senda, H. Izumi, and H. Fujimura, “On uracil deriva- tives and related compounds. 6, 5-alkyl-2,4,6-trioxo-per- hydrophyrimidine derivatives as antiphlogistics,” Arznei- mittelforschung; vol. 17, no.12, December 1967, pp. 1519-1523.
[2] N. Takamura , T. maruyama, E. Chosa, K. Kawai, Y. Tsutsumi, Y. Uryu, K. Yamasaki, T. Deguchi, and M. Otagiri, “Bucolome, a potent binding inhibitor for fu- rosemide, alters the pharmacokinetics and diuretic effect of furosemide: potential for use of bucolome to restore diuretic response in nephrotic syndrome,” Drug Metabo- lism and Disposition,vol. 33, no. 4, April 2005, pp. 596- 602.
[3] M. Osawa, N. Hada, K. Matsumoto, T. Hasegawa, D. Kobayashi, Y. Morimoto, M. Yamaguchi, I. Kanamoto, T. Nakagawa, and K. Sugibayashi, “Usefulness of coadmin- istration of bucolome in warfarin therapy: pharmacoki- netic and pharmacodynamic analysis using outpatient prescriptions,” Internal Journal of Pharmaceutics, vol. 293, no. 1-2, April 2005, pp. 43-49.
[4] M. Kobayashi, M. Takagi, K. Fukumoto, R. Kato, K. Tanaka, and K.Ueno, “The effect of bucolome, a CYP2C9 inhibitor, on the pharmacokinetics of losartan,” Drug Metabolism and Disposition, vol. 23, no. 2, 2008, pp. 115-119.
[5] B. K. Tang, W. Kalow, and A. A. Grey, “Metabolic Fate of Phenobarbital in man N-Glucoside Formation,” Drug Metabolism and Disposition, vol. 5, no. 5, September- October, 1979, pp. 315-318.
[6] B. K. Tang, W. Kalow, and A.A. Grey, “Amobarbital metabolism in man: N-glucoside formation,”Research communications in chemical pathology and pharmacol- ogy, vol. 21, no. 1, July, 1978, 45-53.
[7] S. M. Neighbors, and W. H. Soine, “Identification of Phenobarbital N-glucuronides as urinary metabolites of Phenobarbital in mice,” Drug Metabolism and Disposi- tion, vol. 23, no. 5, May, 1995, pp. 548-552.
[8] K. Mohri, T. Uesugi, and K. Kamisaka, “Bucolome N- glucuronide: purification and identification of a major metabolite of bucolome in rat bile,” Xenobiotica, vol. 15, no. 7, July, 1985, pp. 615-621.
[9] K. Mohri, and Y. Uesawa, “Enzymatic Activities in the Microsomes Prepareted from Rat Small Intestinal Epithelial Cells by Differential Procedures,” Pharmaceutical Research, vol. 18, no. 8, Augasut, 2001, pp. 1232-1236.
[10] O. H. Lowry, N .J. Rosebrough, A. L. Farr, and R. J. Randall, “Protein measurement with the folin phenol reagent,” Journal of Biological Chemistry, vol. 1, no. 1, November, 1951; pp. 265-275.
[11] P. K. Smith, R. I. Krohn, G.T. Hermanson, A. K. Malliam F. H. Garther, M. D. Provenzano, E. K. Fujimoto, N. M. Goeke, B. J. Olson, and D. C. Klenk, “Measurement of protein using bicinchoninic acid,” Analitycal Biochemistry, vol. 150, no. 1, October, 1985, pp. 76-85.
[12] S. Ikushiro, M. Sahara, Y. Emi, Y. Yabusaki, and T. Iya- nagi, “Functional Coexpression of Xenobiotic Metabo- lizing Enzymes, Rat Cytochrome P4501A1 and UDP- Glucuronosyltransferase 1A6, in Yeast Microsomes,” Bio- chimica et Biophysica Acta , vol. 1672, 2004, pp. 86-92.
[13] T. Iyanagi, M. Haniu, K. Sogawa, Y. Fujii-Kuriyama, S. Watanabe, J. E. Shively, and K. F. Anan, “Cloning and characterization of cDNA encoding 3-methylcholanthre- ne inducible rat mRNA for UDP-glucuronosyltransfer- ase,” Journal of Biological Chemistry, vol. 261, no. 33, November, 1986, pp. 15607-15614.
[14] T. Iyanagi, “Molecular basis of multiple UDP-glucurono- syltransferase isoenzyme deficiencies in the hyperbiliru- binemic rat (Gunn rat),” Journal of Biological Chemistry, vol. 266, no. 35, December, 1991, pp. 24048-24052.
[15] Y. Emi, S. Ikushiro, and Y. Kato, “Thyroxine-metabo- lizing Rat UDP-glucuronosyltransferase 1A7 is Regulated by Thyroid Hormone Receptor,” Endocrinology, vol. 148, no. 12, December, 2007, pp. 6124-6133.
[16] H. Ito, Y. Fukuda, K. Murata, and A. Kimura, “Trans- formation of intact yeast cells treated with alkali cations,” Journal of Biological Chemistry, vol. 153, no. 1, January, 1983, pp. 163-168.
[17] K. Oeda, T. Sakai, and H. Ohkawa, “Expression of rat liver cytochrome P-450MC cDNA in Saccharomyces cerevisiae,” DNA, vol. 4, no. 3, June, 1985, pp. 203-210.
[18] S. Ikushiro, Y. Emi, and T. Iyanagi, “Identification and analysis of drug-responsive expression of UDP-glu- curonosyltransferase family 1 (UGT1) isozymes in rat hepatic microsomes using anti-peptide antibodies,” Ar- chives of Biochemistry and Biophysics, vol, 324, no. 2, December, 1995, pp. 267-272.
[19] N. Kasai, T. Sakai, R. Shinkyo, S. Ikushiro, T. Iyanagi, M. Kamao, T, Okano, and K. Inoue, “Sequential Metabolism of 2,3,7-Trichlorodibenzo-p-dioxin (2,3,7-triCDD) by Cytochrome P450 and UDP-Glucuronosyltransferase in Human Liver Microsomes,” Drug Metabolism and Dis- position, vol. 32, no. 8, August, 2004, pp. 870-875.
[20] K. Mohri, Y. Uesawa, and T. Uesugi, “Metabolisme of bucolome in rats. Stability and biliary excretion of buco- lome N-glucuronide,” Journal of Chromatogrphy B, vol. 759, no. 1, August, 2001, pp. 153-159.
[21] N. R. Vansell, and C. D. Klaassen, “Increase in rat liver UDP-glucuronosyltransferase mRNA by microsomal en- zyme inducers that enhance thyroid hormone glucuroni- dation,” Drug Metabolism and Disposition, vol, 30, no.3, March, 2002, pp. 240-246.
[22] M. K. Shelby, N.J. Cherrington, N. R. Vansell, and C. D. Klaassen, “Tissue mRNA Expression of the Rat UDP- Glucuronosyltransferase Gene Family,” Drug Metabolism and Disposition, vol. 31, no. 3, March, 2003, pp. 326- 333.
[23] J. R. Chowdhury, R. Kondapalli, and N. R. Chowdhury, “Gunn rat: A model for inherited deficiency of bilirubin glucuronidation,” Advances in Veterinary Science and Comparative Medicine, vol. 37, 1993, pp. 149-173.
[24] H. Kanoh, K. Okada, and K. Mohri, “Identification of the UDP-glucuronosyltransferase responsible for bucolome N-glucuronide formation in rats,” Pharmazie, vol. 65, no. 11, November, 2010, pp. 840-844.

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